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Biology subjects

Di Giusto, P.

Publications and source records attributed to Di Giusto, P..

3 recordsLinked to original sources

Human Genome-Scale Models of Metabolism and Gene Expression Reveal Resource Constraints of Cancer Cell Lines

Genome-scale metabolic models (M-models) provide mechanistic insight into intracellular metabolism by simulating fluxes subject to nutrient and energy resource constraints. However, they cannot account for a major component of resource allocation, since they do not explicitly account for the cost of producing and maintaining enzymes. Genome-scale models of metabolism and gene expression (ME-Models) address this by including gene expression reactions, but these have only been developed for prokaryotes due to the additional complexity and challenges of modeling eukaryotes. Here, we present the human ME-Model, which encodes transcription, translation, complex formation, and turnover reactions for all enzymes catalyzing metabolic reactions, and couples these processes to constrain metabolic fluxes. We introduce humanME, a Python package to build and analyze human ME-Models. With this, we constructed 16 cancer cell line ME-Models. We found that resource constraints improve growth-rate predictions, and that ME-Model flux predictions are more biologically plausible and efficient. Moreover, transcriptional fluxes recapitulate RNA-Seq expression levels, with discrepancies revealing potential trade-offs involving multiple cellular objectives. Finally, the ME-Model recapitulates the Warburg effect, with increasing growth rate inducing glycolytic shifts, in part due to machinery costs of the electron transport chain. Altogether, we show ME-modeling can mechanistically link gene expression, resource allocation, and metabolism in human cells, substantially expanding the predictive scope of constraint-based models.

systems biology↗

A community-consensus reconstruction of Chinese Hamster metabolism enables structural systems biology analyses to decipher metabolic rewiring in lactate-free CHO cells

Genome-scale metabolic models (GEMs) are indispensable for studying and engineering cellular metabolism. Here, we present iCHO3K, a community-consensus, manually-curated reconstruction of the Chinese Hamster metabolic network. In addition to accounting for 11004 reactions associated with 3597 genes, iCHO3K includes 3489 protein structures and structural descriptors for >70% of its 7377 metabolites, enabling deeper exploration of the link between molecular structure and cellular metabolism. We used iCHO3K to contextualize transcriptomics and metabolomics data from a CHO cell line in which lactate secretion is abolished. We found the reduced glycolytic flux and enhanced TCA cycle flux were accompanied by an elevated NADH and PEP levels in these cells, consistent with experimental measurements. Leveraging iCHO3Ks structural annotations, we identified candidate binding interactions of NADH and PEP with glycolytic enzymes showing model-predicted differential flux, suggesting novel allosteric regulation associated with the observed decrease in glucose uptake and glycolysis. Overall, iCHO3K offers a valuable framework for systematic integration of omics data, improved flux predictions, and structure-guided insights, thus advancing CHO cell engineering and enhancing biomanufacturing efficiency.

systems biology↗

A reconstruction of the mammalian secretory pathway identifies mechanisms regulating antibody production

The secretory pathway processes >30% of mammalian proteins, orchestrating their synthesis, modification, trafficking, and quality control. However, its complexity-- spanning multiple organelles and dependent on coordinated protein interactions--limits our ability to decipher how protein secretion is controlled in biomedical and biotechnological applications. To advance such research, we present secRecon--a comprehensive reconstruction of the mammalian secretory pathway, comprising 1,127 manually curated genes organized within an ontology of 77 secretory process terms, annotated with functional roles, subcellular localization, protein interactions, and complex composition. Using secRecon to integrate multi-omics data, we identified distinct secretory topologies in antibody-producing plasma cells compared to CHO cells. Genes within proteostasis, translocation, and N-glycosylation are deficient in CHO cells, highlighting them as potential engineering targets to boost secretion capacity. Applying secRecon to single-cell transcriptomics and SEC-seq data, we uncovered secretory pathway signatures underlying secretion diversity among IgG-secreting plasma cells. Different transcriptomic clusters had unique secretory phenotypes characterized by variations in the unfolded protein response (UPR), endoplasmic reticulum-associated degradation (ERAD), and vesicle trafficking pathways. Additionally, we discovered specific secretory machinery genes as new markers for plasma cell differentiation. These findings demonstrate secRecon can identify mechanisms regulating protein secretion and guide diverse studies in biomedical research and biotechnology. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=73 SRC="FIGDIR/small/623668v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@f9f865org.highwire.dtl.DTLVardef@122db7forg.highwire.dtl.DTLVardef@1788a1aorg.highwire.dtl.DTLVardef@b2b2bd_HPS_FORMAT_FIGEXP M_FIG C_FIG

systems biology↗